大肠杆菌guaC基因的克隆及高效表达

以E.coli MC4100染色体DNA为模板使用PCR扩增技术克隆了鸟苷酸还原酶基因guaC，该基因全长1044bp，编码相对分子质量为37ku的蛋白质。将该基因直接克隆于PET—16b质粒的T7启动于下游，得到质粒pEXP1。转化E．coli BL21(DE3)，经异丙基硫代半乳糖苷(IPTG)诱导，获得了高效的表达。含表达质粒的菌体胞内粗提液经酶活分析表明，鸟苷酸还原酶的活性为不含质粒的宿主菌的80-85倍。     

人细胞色素P450氧化还原酶的克隆表达和抗体的制备

细胞色素P450还原酶(CPR)是人细胞色素P450酶系的电子供给者,对后者活性的发挥起到重要作用.本研究将从人胚胎cDNA文库中得到的人CPR的编码基因,连接入pT7450表达载体中,在大肠杆菌BL21(DE3)中高效表达并纯化.每升发酵液可得到纯化蛋白49 mg,占总蛋白含量的10%,以细胞色素C为底物检测酶活性,比活为65 u/mg.利用纯化的CPR作为抗原,常规免疫纯系大耳家兔,获得高效价、高特异性的抗血清,抗体滴度为1∶200 000(ELISA法),纯化后抗体应用于不同组织中CPR含量的评价,证明重组CPR及其抗体可用于细胞色素P450的体外药物代谢及细胞色素P450与CPR作用机理的研究.     

Nitric oxide synthase reduces nitrite to NO under anoxia

Cultured bEND.3 endothelial cells show a marked increase in NO production when subjected to anoxia, even though the normal arginine pathway of NO formation is blocked due to absence of oxygen. The rate of anoxic NO production exceeds basal unstimulated NO synthesis in normoxic cells. The anoxic release of NO is mediated by endothelial nitric oxide synthase (eNOS), can be abolished by inhibitors of NOS and is accompanied by consumption of intracellular nitrite. The anoxic NO release is unaffected by the xanthine oxidase inhibitor oxypurinol. The phenomenon is attributed to anoxic reduction of intracellular nitrite by eNOS, and its magnitude and duration suggests that the nitrite reductase activity of eNOS is relevant for fast NO delivery in hypoxic vascular tissues. Received 20 August 2006; received after revision 21 September 2006; accepted 8 November 2006     

硝酸铈铵氧化体系固态合成双偶氮化合物

报道了一种在固态条件下合成二芳基双偶氮化合物的新方法.用硝酸铈铵(CAN)氧化体系对二芳基卡巴肼进行双侧氧化脱氢反应,共合成了8种二芳基卡巴二腙.这种方法具有条件温和、操作简便、无溶剂、污染少等优点.     

All in the family: aldose reductase and closely related aldo-keto reductases

Aldose reductase catalyzes the first step in the polyol pathway and is thought to be involved in the pathogenesis of diabetic complications. In addition to polyol synthesis, aldose reductase may have multiple other activities that intersect with signal processing and oxidative defense mechanisms. Multiple aldose reductase-like proteins have been discovered to have structures and catalytic properties that broadly overlap those of aldose reductase. This chapter will summarize new insights into properties and functions of aldose reductase and closely related members of the aldo-keto reductase enzyme superfamily.     

Functional, biochemical and genetic diversity of prokaryotic nitrate reductases

Prokaryotic nitrate reduction can serve a number of physiological roles and can be catalysed by a number of biochemically distinct nitrate reductases. Three distinct nitrate reductase classes can be indentified in prokaryotes, NAS, NAR and NAP. NAS is located in the cytoplasmic compartment and participates in nitrogen assimilation. NAR is usually a three-subunit complex anchored to the cytoplasmic face of the membrane with its active site located in the cytoplasmic compartment and is involved in anaerobic nitrate respiration. NAP is a two-subunit complex, located in the periplasmic compartment, that is coupled to quinol oxidation via a membrane anchored tetraheme cytochrome. It shows considerable functional flexibility by participating in anaerobic respiration or redox energy dissipation depending on the organism in which it is found. The members of all three classes of enzymes bind the bis-molybdopterin guanine dinucleotide cofactor at the active site, but they differ markedly in the number and nature of cofactors used to transfer electrons to this site. Analysis of prokaryotic genome sequences available at the time of writing reveals that the different nitrate reductases are phylogenetically widespread.     

中国人群MTHFR基因A677V多态性

目的：研究在广东汉族人群中四氢叶酸还原酶基因６７７位点是否存在多态性。方法：运用多聚酶链反应－限制性内切酶片段长度多态性技术（ＰＣＲ－ＲＦＬＰ）检测ＭＴＨＦＲ的６７７位点多态性。结果：在中国广东汉族人群中,Ｃ等位基因的频率为０．７。结论：ＭＴＨＦＲ的６７７位碱基存在着多态性,为进一步研究该位点多态性与妊高征的相关性奠定了基础。     

硝酸还原酶和可溶性蛋白对东北三省蒙古栎种源生长性状的影响

本文研究了东北三省25个蒙古栎种源的硝酸还原酶和可溶性蛋白与生长性状的关系．不同生长性状的蒙古栎种源,硝酸还原酶活性和可溶性蛋白含量上都存在着显著的差异,并且同一种源呈现明显的季节变化．试验结果表明生长性状与所研究的两个理化指标之问存在显著的正相关,即高、径生长好的种源,硝酸还原酶活性和可溶性蛋白含量高;硝酸还原酶活性大的种源,可溶性蛋白含量也高,可以为林木育种提供参考．本试验中,集安种源在两项生理指标中含量均最高,可作为优质种源进行推广．     

An Affordable Accurate Evaluation of the Binding Conformations Predicted by Molecular Docking Studies in Bio-System

正Recently,docking has been widely used to predict the binding-modes of protein-inhibitors,when the crystal complexes structure was absent.Most docking algorithms are able to generate a large number of probable conformations,it,however,is difficult to effectively evaluate these docking poses and identify the most reasonable bindingmode.In the present study,on the basis of the crystallographic data of human 3-hydroxy-3-methylglutaryl coenzyme     

硝酸稀土与速克灵混配对番茄灰霉病的防病效果及机制的研究

室温下,用1500倍速克灵、3000mg/L硝酸稀土、1500倍速克灵+3000mg/L硝酸稀土、2000倍速克灵+3000mg/L硝酸稀土、2500倍速克灵+3000mg/L硝酸稀土、3000倍速克灵+3000mg/L硝酸稀土的混剂喷施番茄幼苗,间隔不同时间接种番茄灰霉病菌,测定其防病效果及番茄叶片内丙二醛含量和多酚氧化酶活性。结果表明:不同处理在喷施后第15天时,对番茄灰霉病的防病效果最为理想,其中1500倍速克灵+3000mg/L硝酸稀土的混剂的效果最好,防效达96.10%,1500倍速克灵、3000mg/L硝酸稀土和2000倍速克灵+3000mg/L硝酸稀土、2500倍速克灵+3000mg/L硝酸稀土、3000倍速克灵+3000mg/L硝酸稀土的混剂次之。喷施1500倍速克灵+3000mg/L硝酸稀土混剂的植株丙二醛含量最低,为30.54(mmol·g),多酚氧化酶活性最高,为73(0.01A/min)。说明稀土与速克灵混合具有良好的抑菌和增抗作用。